To analyze the effect of RNASE2, the cell proliferation rate, level of apoptosis, migration, and invasion capabilities of cells in the shNC, shRNASE2, ovNC, and ovRNASE2 groups were measured. After culturing for 1, 2, 3, and 4 days, the optical density of cells at 450 nm (OD450 nm) was measured according to the method of the Cell Counting Kit-8 (KeyGen, Nanjing, China). The cell proliferation rate was calculated using the OD450 nm in the following equation: Cell proliferation rate = (OD value at other time points/OD value at 0 h − 1) × 100% (same sample). After culturing for 48 h, the level of apoptotic cells was determined using the Annexin V-APC/7-AAD Apoptosis Kit (Abnova Corporation, Taibei, China). Cell migration and invasion were evaluated using Transwell and Transwell-Matrigel assays (22 (link)).
To investigate whether RNASE2 plays a role through the PI3K/AKT pathway, LY294002, a PI3K inhibitor, was used to treat cells in the ovRNASE2 group, termed ovRNASE2+LY294002. The ovNC and ovRNASE2 groups were treated with DMSO for use as controls and were named ovNC+DMSO and ovRNASE2+DMSO, respectively. The cell proliferation rate, level of apoptosis, and migration and invasion capabilities of these groups were measured using the same methods as those described above.
To investigate whether RNASE2 plays a role through the PI3K/AKT pathway, LY294002, a PI3K inhibitor, was used to treat cells in the ovRNASE2 group, termed ovRNASE2+LY294002. The ovNC and ovRNASE2 groups were treated with DMSO for use as controls and were named ovNC+DMSO and ovRNASE2+DMSO, respectively. The cell proliferation rate, level of apoptosis, and migration and invasion capabilities of these groups were measured using the same methods as those described above.
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